Elucidating temporal trends in trace element exposure of green turtles (Chelonia mydas) using the toxicokinetic differences of blood and scute samples.

Blood is considered a suitable biomonitoring matrix for evaluating relatively recent exposure to environmental contaminants since abrupt changes in exposure regimes are rapidly reflected in blood. On the other hand, keratinized tissues, such as turtle scutes, are known to integrate trace element exposure over relatively long time periods. This study aimed to test the use of the differences in blood and scute to inform on the historical trace element exposure of green turtles. We propose a blood-scute kinetic model to predict how an increase in exposure would affect the concentrations in these two matrices over time. We then tested the relationship between blood and scute concentrations for 19 trace elements in two green turtle populations presumed to experience relatively constant exposure conditions. Significant log-log and linear correlations were observed between blood and scute concentrations for Co, As, Mo, Sb, and Cd. We then analysed blood-scute ratios in turtles from two coastal sites with known elevated exposure to various trace elements from previous studies. Deviations from the steady-state were clearly evident in these coastal turtles (for Co and Cd) and were consistent with the model prediction of changes in exposure. These field data provide evidence that blood-scute ratios can provide a valuable tool for examining the historical trace element exposure of turtles. We further present a method by which the general model may be refined and validated, by using data from individual turtles that had been recaptured across multiple years. Although the timeframe and number of recaptured samples available for this study were limited, the temporal changes in blood-scute ratios in these animals were generally consistent with those suggested by the model. Thus, the ratio between paired blood and scute trace element concentrations could be used to establish a temporal exposure index in turtles.

Trace element reference intervals in the blood of healthy green sea turtles to evaluate exposure of coastal populations.

Exposure to essential and non-essential elements may be elevated for green sea turtles (Chelonia mydas) that forage close to shore. Biomonitoring of trace elements in turtle blood can identify temporal trends over repeated sampling events, but any interpretation of potential health risks due to an elevated exposure first requires a comparison against a baseline. This study aims to use clinical reference interval (RI) methods to produce exposure baseline limits for essential and non-essential elements (Na, Mg, K, Ca, Ti, V, Cr, Mn, Fe, Co, Ni, Cu, Zn, As, Se, Mo, Cd, Sb, Ba, and Pb) using blood from healthy subadult turtles foraging in a remote and offshore part of the Great Barrier Reef. Subsequent blood biomonitoring of three additional coastal populations, which forage in areas dominated by agricultural, urban and military activities, showed clear habitat-specific differences in blood metal profiles relative to the those observed in the offshore population. Coastal turtles were most often found to have elevated concentrations of Co, Mo, Mn, Mg, Na, As, Sb, and Pb relative to the corresponding RIs. In particular, blood from turtles from the agricultural site had Co concentrations ranging from 160 to 840 µg/L (4-25 times above RI), which are within the order expected to elicit acute effects in many vertebrates. Additional clinical blood biochemistry and haematology results indicate signs of a systemic disease and the prevalence of an active inflammatory response in a high proportion (44%) of turtles from the agricultural site. Elevated Co, Sb, and Mn in the blood of these turtles significantly correlated with elevated markers of acute inflammation (total white cell counts) and liver dysfunction (alkaline phosphatase and total bilirubin). The results of this study support the notion that elevated trace element exposures may be adversely affecting the health of nearshore green sea turtles.

A decision framework for prioritizing multiple management actions for threatened marine megafauna.

Resources for conserving biodiversity are invariably insufficient. This situation creates the need for transparent, systematic frameworks to help stakeholders prioritize the allocation of resources across multiple management actions. We developed a novel framework that explicitly prioritizes actions to minimize the impacts of several threats across a species' range. The framework uses a budget constraint and maximizes conservation outcomes from a set of management actions, accounting for the likelihood of the action being successfully applied and accepted by local and Indigenous communities. This approach is novel in that it integrates local knowledge and expert opinion with optimization software, thereby minimizing assumptions about likelihood of success of actions and their effectiveness. To test the framework, we used the eastern Gulf of Carpentaria and Torres Strait population of the flatback turtle, Natator depressus, as a case study. This approach allowed the framework to be applied in a data-poor context, a situation common in conservation planning. The framework identified the best set of actions to maximize the conservation of flatback eggs for scenarios with different budgets and management parameters and allowed comparisons between optimized and preselected scenarios. Optimized scenarios considered all implementable actions to explore how to best allocate resources with a specified budget and focus. Preselected scenarios were used to evaluate current allocations of funds and/or potential budget allocations suggested by different stakeholders. Scenarios that used a combination of aerial and ground strategies to reduce predation of eggs performed better than scenarios that focused only on reducing harvest of eggs. The performances of optimized and preselected scenarios were generally similar among scenarios that targeted similar threats. However, the cost-effectiveness of optimized scenarios was usually higher than that of preselected scenarios, demonstrating the value of conducting a systematic optimization approach. Our method provides a foundation for more effective conservation investments and guidance to prioritize actions within recovery plans while considering the sociopolitical and cultural context of decisions. The framework can be adapted easily to a wide range of species, geographical scales, and life stages.

A risk assessment approach to manage inundation of Elseya albagula nests in impounded waters: a win-win situation?

A risk assessment process was used to trial the impact of potential new operating rules on the frequency of nest inundation for the White-throated snapping turtle, Elseya albagula, in the impounded waters of the Burnett River, Queensland, Australia. The proposed operating rules would increase the barrage storage level during the turtle nesting season (May-July) and then would be allowed to reduce to a lower level for incubation for the rest of the year. These proposed operating rules reduce rates of nest inundation by altering water levels in the Ben Anderson Barrage impoundment of the Burnett River. The rules operate throughout the turtle reproductive period and concomitantly improve stability of littoral habitat and fishway operation. Additionally, the proposed rules are expected to have positive socio-economic benefits within the region. While regulated water resources will inherently have a number of negative environmental implications, these potential new operating rules have the capacity to benefit the environment while managing resources in a more sustainable manner. The operating rules have now been enacted in subordinate legislation and require the operator to maintain water levels to minimize turtle nest inundation.

Plasma biochemical and PCV ranges for healthy, wild, immature hawksbill (Eretmochelys imbricata) sea turtles.

In recent years, the use of blood chemistry as a diagnostic tool for sea turtles has been demonstrated, but much of its effectiveness relies on reference intervals. The first comprehensive blood chemistry values for healthy wild hawksbill (Eretmochelys imbricata) sea turtles are presented. Nineteen blood chemistry analytes and packed cell volume were analysed for 40 clinically healthy juvenile hawksbill sea turtles captured from a rocky reef habitat in northern Australia. We used four statistical approaches to calculate reference intervals and to investigate their use with non-normal distributions and small sample sizes, and to compare upper and lower limits between methods. Eleven analytes were correlated with curved carapace length indicating that body size should be considered when designing future studies and interpreting analyte values.

Corneal fibropapillomatosis in green sea turtles (Chelonia mydas) in Australia.

Chelonid corneal fibropapillomatosis has not previously been recorded in Australian waters. During 2008, 724 green sea turtles (Chelonia mydas) were examined in Queensland, Australia at two sites, Moreton Bay (n=155) and Shoalwater Bay (n=569), during annual monitoring. In the same calendar year, 63 turtles were submitted from various sites in southern Queensland for post-mortem examination at the University of Queensland. Four of the 787 animals (0.5%) were found to have corneal fibropapillomas of varying size, with similar gross and microscopical features to those reported in other parts of the world. Two animals with corneal fibropapillomas also had cutaneous fibropapillomas. Clinical assessment indicated that these lesions had detrimental effects on the vision of the turtles and therefore their potential ability to source food, avoid predators and interact with conspecifics. Importantly, these findings represent an emergence of this manifestation of fibropapillomatosis in green sea turtle populations in the southern Pacific Ocean.

Evidence for transoceanic migrations by loggerhead sea turtles in the southern Pacific Ocean.

Post-hatchling loggerhead turtles (Caretta caretta) in the northern Pacific and northern Atlantic Oceans undertake transoceanic developmental migrations. Similar migratory behaviour is hypothesized in the South Pacific Ocean as post-hatchling loggerhead turtles are observed in Peruvian fisheries, yet no loggerhead rookeries occur along the coast of South America. This hypothesis was supported by analyses of the size-class distribution of 123 post-hatchling turtles in the South Pacific and genetic analysis of mtDNA haplotypes of 103 nesting females in the southwest Pacific, 19 post-hatchlings stranded on the southeastern Australian beaches and 22 post-hatchlings caught by Peruvian longline fisheries. Only two haplotypes (CCP1 93% and CCP5 7%) were observed across all samples, and there were no significant differences in haplotype frequencies between the southwest Pacific rookeries and the post-hatchlings. By contrast, the predominant CCP1 haplotype is rarely observed in North Pacific rookeries and haplotype frequencies were strongly differentiated between the two regions (F(st)=0.82; p=<0.00001). These results suggest that post-hatchling loggerhead turtles emerging from the southwest Pacific rookeries are undertaking transoceanic migrations to the southeastern Pacific Ocean, thus emphasizing the need for a broader focus on juvenile mortality throughout the South Pacific to develop effective conservation strategies.

Flame retardants (PBDEs) in marine turtles, dugongs and seafood from Queensland, Australia.

Polybrominated diphenyl ethers (PBDEs) are used as flame retardants in numerous products. These compounds have been found to enter the marine environment where they have the potential to bioaccumulate in biota. Limited information is currently available concerning the levels of PBDEs in Australian marine wildlife. This study presents baseline information on PBDE levels in a variety of marine species from Queensland, Australia and considers the influence of species-specific factors on contaminant levels and tissue distribution in marine turtles. Overall, the PBDE levels measured in this study are relatively low compared to marine biota from the northern hemisphere, indicating low level input into the marine system of Queensland. This is in general agreement with global estimates which suggest low PBDE usage in Australia. Previous studies, however, have found relatively high PBDE levels in Australian human milk and sera. This discrepancy in contamination trends between terrestrial and marine biota suggests that future transport of PBDEs may occur to the marine system in Australia.

Patterns of lipid storage and mobilisation in the female green sea turtle ( Chelonia mydas).

Reproductive data from southern Queensland indicate that vitellogenesis in female Chelonia mydas takes approximately 8 months and is followed by a migration to a breeding area. At Heron Island, females lay multiple clutches over approximately 3 months. To investigate how females mobilise and store lipid during the breeding season we collected plasma, yolk, and fat tissue samples from females at a variety of stages during the nesting season. In breeding females, concentrations of plasma triglyceride increased seasonally. They reached peak concentrations during vitellogenesis and courtship, remained high throughout the nesting season, and then declined to a nadir after the last clutch. Plasma protein concentration increased throughout the breeding season, peaking following the last clutch for the season. Yolk lipids were highest during courtship and were similar throughout the nesting season, suggesting that uptake of lipid by ovarian follicles is completed prior to the beginning of the nesting season. Plasma triglyceride decreases in females with prolonged periods of unsuccessful nesting, and total lipid levels in adipose tissue and follicle yolks were significantly lower in atretic females. It appears that: (1) endogenous energy reserves can be reduced by stochastic environmental events (such as those reducing nesting success), and (2) a metabolic shift signalling the end of the nesting season is characterised by a drop in plasma triglycerides and slight increase in total plasma protein.

Quantitative analysis of herpesvirus sequences from normal tissue and fibropapillomas of marine turtles with real-time PCR.

Quantitative real-time PCR has been used to measure fibropapilloma-associated turtle herpesvirus (FPTHV) pol DNA loads in fibropapillomas, fibromas, and uninvolved tissues of green, loggerhead, and olive ridley turtles from Hawaii, Florida, Costa Rica, Australia, Mexico, and the West Indies. The viral DNA loads from tumors obtained from terminal animals were relatively homogeneous (range 2-20 copies/cell), whereas DNA copy numbers from biopsied tumors and skin of otherwise healthy turtles displayed a wide variation (range 0.001-170 copies/cell) and may reflect the stage of tumor development. FPTHV DNA loads in tumors were 2.5-4.5 logs higher than in uninvolved skin from the same animal regardless of geographic location, further implying a role for FPTHV in the etiology of fibropapillomatosis. Although FPTHV pol sequences amplified from tumors are highly related to each other, single signature amino acid substitutions distinguish the Australia/Hawaii, Mexico/Costa Rica, and Florida/Caribbean groups.

Evidence for a hormonal tactic maximizing green turtle reproduction in response to a pervasive ecological stressor.

Mortality of breeding sea turtles due to excessive heat exposure after nesting activities is an unusual feature of the Raine Island green turtle rookery. Breeding turtles that fail to return to the ocean after oviposition can experience increasing body temperatures that exceed lethal limits (>39 degrees C) as ambient temperatures rise after sunrise. We investigated how acute increases in body temperature influenced plasma corticosterone (B) concentrations of individual turtles. Furthermore, interactions between progesterone (P) and testosterone (T) and increasing body temperature and the glucocorticoid corticosterone were examined for negative correlations. Breeding green turtles exhibited a 16-fold mean increase in plasma corticosterone concentration as body temperature (cloacal) rose from 28.2 to 40.7 degrees C in less than 6 h. However, the absolute increase in plasma B was small and much less than expected, despite the lethal stressor. Comparatively, the maximal B response to lethal heat stress was similar to plasma B concentrations obtained from breeding female turtles exposed to 8 h of capture stress. However, the maximal B response of breeding turtles exposed to heat and capture stressors was significantly less than the B response of nonbreeding adult female turtles subjected to an 8-h capture stressor. No negative correlations were observed between plasma T and plasma B, between plasma T and body temperature, between plasma P and plasma B, or between plasma P and body temperature. Our findings provide further evidence that reduced adrenocortical function operates in breeding green turtles in the presence of even the most pervasive of environmental stressors.

Interactions between behavior and plasma steroids within the scramble mating system of the promiscuous green turtle, Chelonia mydas.

We measured plasma androgen (combined testosterone and 5alpha-dihydrotestosterone) (A) and corticosterone (B) in the promiscuous green turtle (Chelonia mydas) during courtship in the southern Great Barrier Reef. This study examined if reproductive behaviors and intermale aggression induced behavioral androgen and adrenocortical responses in reproductively active male and female green turtles. Associations between reproductive behavior and plasma steroids were investigated in green turtles across the population and within individuals. Levels across a range of both asocial and social behaviors were compared including (a) free swimming behavior; (b) initial courtship interactions; (c) mounted behavior (male and female turtles involved in copulatory activities); (d) intermale aggression (rival males that physically competed with another male turtle or mounted males recipient to these aggressive interactions); and (e) extensive courtship damage (male turtles that had accumulated excessive courtship damage from rival males). Behavioral androgen responses were detected in male turtles, in that plasma A was observed to increase with both attendant and mounted behavior. Male turtles who had been subjected to intermale aggression or who had accumulated severe courtship damage exhibited significantly lower plasma A than their respective controls. No pronounced adrenocortical response was observed after either intermale aggression or accumulation of extensive courtship damage. Female turtles exhibited a significant increase in plasma B during swimming versus mounted behavior, but no change in plasma A. We discuss our results in terms of how scramble polygamy might influence behavioral androgen interactions differently from more typical combative and territorial forms of male polygamy.

Plasma steroid interactions during high-density green turtle nesting and associated disturbance.

Raine Island in the Northern Great Barrier Reef constitutes an extremely high-density green turtle (Chelonia mydas) rookery. On this island, competitive interactions for nesting space and subsequent disturbance of individual nesting are widespread. High-density nesting often delays successful oviposition by one or more nights. There is little information on how hormones in female reptiles interact during competitive reproductive events in such high-density nesting populations. In this three-part study we investigated the interactions between density (within and between rookery/ies), nesting success and failure, and plasma steroid profiles in green turtles. First, we compared levels of plasma corticosterone (B) and combined testosterone + 5alpha-dihydrotestosterone (T + DHT) in turtles during five stages of oviposition in both a high-nesting-density sector (1 turtle/m2) and a low-nesting-density sector (0.1 turtle/m2). Second, we investigated the relationship between increasing delays (0, 1, 2, 3, and 6 days) in successful oviposition and the plasma steroids B and T + DHT. Third, we assessed a comparative measure of steroid hormone levels of females at low-density sites on Raine Island (high-density rookery) and Number Seven Sandbank (low-density rookery). Despite a significant trend suggesting high-density nesting turtles elaborated more plasma B than turtles in low-density sectors, the magnitude of this increase was small. We suggest that this increase may be an artifact of increased metabolic demand and hence catabolism of energy substrates associated with high-density nesting. Plasma T + DHT remained stable in response to density-dependent effects associated with nesting. Furthermore, prolonging successful oviposition because of multiple nightly disturbance failed to elicit any change in either plasma B or T + DHT. These data suggest that green turtles may be exhibiting adrenal desensitization to prevent both physical and behavioral disturbances interfering with reproduction. We suspect that down-regulating the acute adrenocortical response may represent an adaptive trade-off mechanism for optimizing current reproductive success at the potential expense of survivorship.

The environmental contaminant DDE fails to influence the outcome of sexual differentiation in the marine turtle Chelonia mydas.

In many turtles, the temperature experienced during the middle of egg incubation determines the sex of the offspring. The implication of steroid sex hormones as the proximate trigger for sex determination opens the possibility that endocrine-disrupting contaminants may also influence the outcome of sexual differentiation. In this study we investigate the potential effects of DDE (a common DDT metabolite) on sexual differentiation of Chelonia mydas (green sea turtle). Four clutches of eggs collected from Heron Island, Queensland, Australia, were treated with DDE at the beginning of the thermosensitive period for sexual determination. An incubation temperature of 28 degrees C or less produces male hatchlings in this species, whereas 30 degrees C or more produces female hatchlings. Dosed eggs were consequently incubated at two temperatures (27.6 degrees C and 30.4 degrees C) on the upper and lower boundaries of the sex determination threshold for this species. DDE, ranging from 3.3 to 66.5 microg, was dissolved in 5, 10, and 25 microl ethanol and applied to eggshells above the embryo. Less than 2.5 ng/g DDE was present in eggs prior to dosing. Approximately 34% of the applied DDE was absorbed in the eggs, but only approximately 8% of applied DDE was found in embryos. Thus, treated eggs, corrected for background DDE, had up to 543 ng/g DDE. The sex ratio at these doses did not differ from what would be expected on consideration of temperature alone. Incubation time, hatching success, incidence of body deformities, hatching size, and weight were also within the limits of healthy developed hatchlings. This indicates that the eggs of C. mydas in the wild with concentrations of DDE less than 543 ng/g should produce hatchlings with relatively high hatching success, survival rate, and normally differentiated gonads.

Geographic structure of mitochondrial and nuclear gene polymorphisms in Australian green turtle populations and male-biased gene flow.

The genetic structure of green turtle (Chelonia mydas) rookeries located around the Australian coast was assessed by (1) comparing the structure found within and among geographic regions, (2) comparing microsatellite loci vs. restriction fragment length polymorphism analyses of anonymous single copy nuclear DNA (ascnDNA) loci, and (3) comparing the structure found at nuclear DNA markers to that of previously analyzed mitochondrial (mtDNA) control region sequences. Significant genetic structure was observed over all regions at both sets of nuclear markers, though the microsatellite data provided greater resolution in identifying significant genetic differences in pairwise tests between regions. Inferences about population structure and migration rates from the microsatellite data varied depending on whether statistics were based on the stepwise mutation or infinite allele model, with the latter being more congruent with geography. Estimated rates of gene flow were generally higher than expected for nuclear DNA (nDNA) in comparison to mtDNA, and this difference was most pronounced in comparisons between the northern and southern Great Barrier Reef (GBR). The genetic data combined with results from physical tagging studies indicate that the lack of nuclear gene divergence through the GBR is likely due to the migration of sGBR turtles through the courtship area of the nGBR population, rather than male-biased dispersal. This example highlights the value of combining comparative studies of molecular variation with ecological data to infer population processes.

Philopatry of male marine turtles inferred from mitochondrial DNA markers.

Recent studies of mitochondrial DNA (mtDNA) variation among marine turtle populations are consistent with the hypothesis that females return to beaches in their natal region to nest as adults. In contrast, less is known about breeding migrations of male marine turtles and whether they too are philopatric to natal regions. Studies of geographic structuring of restriction fragment and microsatellite polymorphisms at anonymous nuclear loci in green turtle (Chelonia mydas) populations indicate that nuclear gene flow is higher than estimates from mtDNA analyses. Regional populations from the northern and southern Great Barrier Reef were distinct for mtDNA but indistinguishable at nuclear loci, whereas the Gulf of Carpentaria (northern Australia) population was distinct for both types of marker. To assess whether this result was due to reduced philopatry of males across the Great Barrier Reef, we determined the mtDNA haplotypes of breeding males at courtship areas for comparison with breeding females from the same three locations. We used a PCR-restriction fragment length polymorphism approach to determine control region haplotypes and designed mismatch primers for the identification of specific haplotypes. The mtDNA haplotype frequencies were not significantly different between males and females at any of the three areas and estimates of Fst among the regions were similar for males and females (Fst = 0.78 and 0.73, respectively). We conclude that breeding males, like females, are philopatric to courtship areas within their natal region. Nuclear gene flow between populations is most likely occurring through matings during migrations of both males and females through nonnatal courtship areas.

Trans-Pacific migrations of the loggerhead turtle (Caretta caretta) demonstrated with mitochondrial DNA markers.

Juvenile loggerhead turtles (Caretta caretta) have recently been documented in the vicinity of Baja California and thousands of these animals have been captured in oceanic fisheries of the North Pacific. The presence of loggerhead turtles in the central and eastern North Pacific is a prominent enigma in marine turtle distribution because the nearest documented nesting concentrations for this species are in Australia and Japan, over 10,000 km from Baja California. To determine the origin of the Baja California feeding aggregate and North Pacific fishery mortalities, samples from nesting areas and pelagic feeding aggregates were compared with genetic markers derived from mtDNA control region sequences. Overall, 57 of 60 pelagic samples (95%) match haplotypes seen only in Japanese nesting areas, implicating Japan as the primary source of turtles in the North Pacific Current and around Baja California. Australian nesting colonies may contribute the remaining 5% of these pelagic feeding aggregates. Juvenile loggerhead turtles apparently traverse the entire Pacific Ocean, approximately one-third of the planet, in the course of developmental migrations, but mortality in high-seas fisheries raises concern over the future of this migratory population.

Mitochondrial DNA control region polymorphisms: genetic markers for ecological studies of marine turtles.

We describe a rapid and sensitive method for the detection of population-specific genetic markers in mitochondrial DNA (mtDNA) and the use of such markers to analyse population structure of marine turtles. A series of oligonucleotide primers specific for the amplification of the mtDNA control region in Cheloniid turtles were designed from preliminary sequence data. Using two of these primers, a 384-385-bp sequence was amplified from the 5' portion of the mtDNA control region of 15 green turtles Chelonia mydas from 12 different Indo-Pacific rookeries. Fourteen of the 15 individuals, including some with identical whole-genome restriction fragment patterns, had sequences that differed by one or more base substitutions. Analysis of sequence variation among individuals identified a total of 41 nucleotide substitutions and a 1-bp insertion/deletion. Comparison with evidence from whole-genome restriction enzyme analysis of the same individuals indicated that this portion of the control region is evolving approximately eight times faster than the average rate and that the sequence analysis detected approximately one fifth of the total variation present in the genome. Restriction enzyme analysis of amplified products from an additional 256 individuals revealed significant geographic structuring in the distribution of mtDNA genotypes among five of the 10 rookeries surveyed extensively. Additional geographic structuring of genotypes was identified through denaturing gradient gel electrophoresis (DGGE) of amplified products. Only two of the 10 rookeries surveyed could not be differentiated, indicating that the Indo-Pacific C. mydas include a number of genetically differentiated populations, with minimal female-mediated gene flow among them. Important applications for genetic markers in the conservation and management of marine turtles include the identification of appropriate demographic units for research and management (i.e. genetically discrete populations) and assessment of the composition of feeding and harvested populations.

Seasonal changes in serum gonadal steroids associated with migration, mating, and nesting in the loggerhead sea turtle (Caretta caretta).

Adult male loggerhead sea turtles, Caretta caretta, exhibited a "prenuptial" spermatogenic cycle that was coincident with increased concentrations of serum testosterone (T). Serum T was high during the months when migration and mating have been recorded for males. In contrast to females, males appear to be annual breeders. Nine reproductively active female C. caretta (as verified through laparoscopy) were tagged with sonic transmitters and were repeatedly bled prior to migration. Four months prior to the nesting season, the ovaries of reproductively active females had hundreds of vitellogenic follicles of approximately 1.5 cm in diameter (i.e., half the size of ovulatory follicles). Approximately 4-6 weeks prior to migration from feeding grounds to mating and nesting areas, serum estradiol-17 beta (E2) concentrations increased significantly and remained high for approximately 4 weeks, suggesting a period of increased vitellogenesis. During a 1- to 2-week period prior to migration, serum E2 decreased significantly, while serum T concentrations increased (at least) until the time of migration. Serum T, E2, and progesterone (PRO) were elevated during nesting if a turtle was going to nest again during that nesting season. During the last nesting of a season, turtles had low serum concentrations of T, E2, and Pro. The prenuptial pattern of gonadal recrudescence and gonadal steroid production in both male and female C. caretta contrasts with those of many temperate freshwater turtles, and this type of reproductive pattern may have been facilitated by adaptation to a tropical marine environment.

An evolutionary link for developing mammalian lungs.

Lungs of the human infant and those of other mammals are filled with fluid immediately prior to birth. Studies of the ionic composition of this fluid indicate that active ionic transport processes occur in the epithelial cells of the potential airspaces. The purpose of this study was to see if these active ion pumps were present in developing species other than mammals thus providing a possible evolutionary link to mammals. A series of samples of lung liquid, amniotic fluid, and plasma were taken from embryonic marine turtles gathered from clutches incubating in the beach at Mon Repos, Queensland, Australia during the summer of 1986-87. The concentrations of sodium, potassium and chloride ions and protein measured in these liquids indicated that active pumping processes similar to that seen in the mammalian lung were present in the developing lungs of these marine reptiles and further, circumstantial evidence was gathered to suggest that this liquid was partially reabsorbed prior to hatching. The results support the notion that processes responsible for the normal development of the human lung and lungs of other mammals are also present in the hollow lungs of marine turtles. Thus there is an evolutionary counterpart controlling lung development in more ancient species. It may be possible to generalize this observation to the development of hollow lungs of other species.